Takahashi Junko, Rudsander Ulla J, Hedenström Mattias, Banasiak Alicja, Harholt Jesper, Amelot Nicolas, Immerzeel Peter, Ryden Peter, Endo Satoshi, Ibatullin Farid M, Brumer Harry, del Campillo Elena, Master Emma R, Scheller Henrik Vibe, Sundberg Björn, Teeri Tuula T, Mellerowicz Ewa J
Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, SLU, Umeå, Sweden.
Plant Cell Physiol. 2009 Jun;50(6):1099-115. doi: 10.1093/pcp/pcp062. Epub 2009 Apr 27.
KORRIGAN1 (KOR1) is a membrane-bound cellulase implicated in cellulose biosynthesis. PttCel9A1 from hybrid aspen (Populus tremula L. x tremuloides Michx.) has high sequence similarity to KOR1 and we demonstrate here that it complements kor1-1 mutants, indicating that it is a KOR1 ortholog. We investigated the function of PttCel9A1/KOR1 in Arabidopsis secondary growth using transgenic lines expressing 35S::PttCel9A1 and the KOR1 mutant line irx2-2. The presence of elevated levels of PttCel9A1/KOR1 in secondary walls of 35S::PttCel9A1 lines was confirmed by in muro visualization of cellulase activity. Compared with the wild type, 35S::PttCel9A1 lines had higher trifluoroacetic acid (TFA)-hydrolyzable glucan contents, similar Updegraff cellulose contents and lower cellulose crystallinity indices, as determined by (13)C solid-state nuclear magnetic resonance (NMR) spectroscopy. irx2-2 mutants had wild-type TFA-hydrolyzable glucan contents, but reduced Updegraff cellulose contents and higher than wild-type cellulose crystallinity indices. The data support the hypothesis that PttCel9A1/KOR1 activity is present in cell walls, where it facilitates cellulose biosynthesis in a way that increases the amount of non-crystalline cellulose.
KORRIGAN1(KOR1)是一种与纤维素生物合成有关的膜结合纤维素酶。来自杂种山杨(Populus tremula L. x tremuloides Michx.)的PttCel9A1与KOR1具有高度的序列相似性,我们在此证明它能互补kor1-1突变体,表明它是KOR1的直系同源物。我们使用表达35S::PttCel9A1的转基因系和KOR1突变体系irx2-2研究了PttCel9A1/KOR1在拟南芥次生生长中的功能。通过在细胞壁内可视化纤维素酶活性,证实了35S::PttCel9A1系次生壁中PttCel9A1/KOR1水平升高。与野生型相比,通过(13)C固态核磁共振(NMR)光谱测定,35S::PttCel9A1系具有更高的三氟乙酸(TFA)可水解葡聚糖含量、相似的Updegraff纤维素含量和更低的纤维素结晶度指数。irx2-2突变体具有野生型的TFA可水解葡聚糖含量,但Updegraff纤维素含量降低且纤维素结晶度指数高于野生型。这些数据支持了这样的假设,即PttCel9A1/KOR1活性存在于细胞壁中,它以增加非结晶纤维素量的方式促进纤维素生物合成。